AIDS is the number-one killer of African-American women between the ages of 25 and 34 in the United States. This group forms the fastest-growing cluster representing 67 percent of all ADDS cases in US. Currently, a vaccine for HIV/AIDS is not available. Since >90% of HIV transmission takes place at the mucosal surfaces, a vaccine that can elicit a potent mucosal immune response is the need of the hour. Currently, non-infectious DNA molecules are being tested but these molecules are weak sources of antigens and do not elicit a strong immune response at mucosal surfaces. Therefore, the goal of this project is to evaluate flagellin to potentiate a plasmid DNA that expresses env, gag and pol genes of HIV. The central hypothesis is that, since flagellin enhances both innate and adaptive immunity, it can potentiate immunogenicity of this DNA at cervix and vagina. The approach is to use this non-infectious HIV-DNA along with flagellin at cervix and vagina of B ALB/c mice. After two intra-vaginal applications of this DNAflagellin complex within 21 days interval, both innate and adaptive immunological responses will be evaluated. The NK cell and dendritic cell activities, and vaccine stimulated cytokine production will be analyzed. In our previous studies, this plasmid DNA has been shown to be immunogenic in muscles. However, this DNA has not been tested at mucosal surfaces. The central hypothesis will be tested and the objective of the project will be accomplished by two specific aims. Aim 1 will test the hypothesis that HIVDNA will be able to generate a strong specific immune response at the reproductive mucosa. Aim 2 will test the adjuvant effect of flagellin at reproductive mucosa. We anticipate that use of an immunogenic plasmid DNA along with a potent adjuvant will mount a strong immune response that will destroy HIV particles present in semen. Since flagellin also activates NK cells and dendritic cells non-specifically, we expected that it will help to destroy cell associated viruses in semen and therefore will assist specific immune responses to prevent virus transmission. It is our expectation that the resultant strategy may prove to be helpful in developing a prophylactic mucosal vaccine for HIV/AIDS thereby reducing chances of virus spread from infected to healthy individuals. Such outcomes will have profound and significant impact on preventing health disparities among minority individuals infected with HIV by providing affordable vaccine and reducing morbidity and mortality associated with HIV and AIDS.